AMPK inhibits CRTC2-mediated hepatic gluconeogenic gene expression via induction of SHP.A, Western blot analysis showing the effects of AICAR on phosphorylation of AMPK and induction of SHP in rat primary hepatocytes treated with 1 mm of AICAR for the indicated times. B, primary rat hepatocytes were infected with Ad-siSHP and constitutively active CRTC2 (Ad-S171A) for 48 h followed by AICAR treatment and harvested for Western blot analysis using the indicated antibodies. C, HepG2 cells were transfected with pSUPER human SHP siRNA expression vector and, after 24 h, subsequently co-transfected with 200 ng of PEPCK-luc, G6Pase-luc, and pcDNA3-FLAG-CRTC2 (S171A) (100 ng) for 24 h and then treated with 1 mm AICAR for 12 h. D, RT-PCR analysis showing the effects of AMPK through SHP expression on mRNA levels of PEPCK and G6Pase in rat primary hepatocytes. Twenty-four hours after the knockdown of SHP using Ad-siSHP in the cells, infected with Ad-CRTC2 (S171A) and Ad-AMPK, total RNA was isolated for quantitative PCR analysis of SHP mRNA expression and was normalized to β-actin expression.